Linking Genetics of Brain Changes to Alzheimer's Disease: Sparse Whole Genome Association Scan of Regional MRI Volumes in the ADNI and AddNeuroMed Cohorts.
Alzheimer's disease (AD) is a highly heritable disease, but until recently few replicated genetic markers have been identified. Markers identified so far are likely to account for only a tiny fraction of the heritability of AD and many more genetic risk alleles are thought to be undiscovered. Identifying genetic markers for AD using combined analysis of genetics and brain imaging data. Imaging quantitative trait loci (iQTLs) has recently emerged as an interesting research area for linking genetics of brain changes to AD. We consider a genome-wide association scan of 109 brain-wide regional imaging phenotypes to identify genetic susceptibility loci for AD from a combined set of 1,045 subjects from the Alzheimer's Disease Neuroimaging Initiative (ADNI) and the AddNeuroMed studies. We use one-SNP-at-a-time as well as multi-SNP Hyperlasso based iQTL methods for the analysis. We identified several novel markers associated with AD, namely HOMER2 (rs1256429; intronic, p = 8.7 × 10⁻¹⁰), EOMES (rs2724509; flanking), JAM2 (rs2829841; intronic), and WEE1 (rs10770042; coding). The SNP rs1256429 (HOMER2) was one of the top hits in Hyperlasso as well as in the single-SNP analysis showing an association with the volume of the right thalamus and AD, a brain region reported to be linked with AD in several studies. We believe that the markers identified in this study are novel additions to the existing list of genetic variants associated with AD which can be validated in future replicated studies.


Monoclonal antibodies to hypothalamic growth hormone-releasing factor with picomoles of antigen.
Monoclonal antibodies specific for rat hypothalamic growth hormone-releasing factor (rGRF) have been produced by in vitro immunization of mouse spleen cells with less than 1 nanomole of rGRF in a partially purified preparation. Hybridoma supernatants were screened for anti-rGRF activity by use of a pituitary culture assay system that can detect growth hormone-releasing factor in the femtomole range. Such highly sensitive in vitro techniques permit the use of picomole quantities of an antigen in partially purified preparations for the isolation of monoclonal antibodies, which can in turn be used in biological studies and in immunochemical procedures for large-scale purification and isolation of that antigen.


Video-rate 3D imaging of living cells using Fourier view-channel-depth light field microscopy.
Interrogation of subcellular biological dynamics occurring in a living cell often requires noninvasive imaging of the fragile cell with high spatiotemporal resolution across all three dimensions. It thereby poses big challenges to modern fluorescence microscopy implementations because the limited photon budget in a live-cell imaging task makes the achievable performance of conventional microscopy approaches compromise between their spatial resolution, volumetric imaging speed, and phototoxicity. Here, we incorporate a two-stage view-channel-depth (VCD) deep-learning reconstruction strategy with a Fourier light-field microscope based on diffractive optical element to realize fast 3D super-resolution reconstructions of intracellular dynamics from single diffraction-limited 2D light-filed measurements. This VCD-enabled Fourier light-filed imaging approach (F-VCD), achieves video-rate (50 volumes per second) 3D imaging of intracellular dynamics at a high spatiotemporal resolution of ~180 nm × 180 nm × 400 nm and strong noise-resistant capability, with which light field images with a signal-to-noise ratio (SNR) down to -1.62 dB could be well reconstructed. With this approach, we successfully demonstrate the 4D imaging of intracellular organelle dynamics, e.g., mitochondria fission and fusion, with ~5000 times of observation.


The energetic cost of short flights in birds.
Many small birds perform short flights, for which take-offs, ascents and descents form a large component of the total flight time and which are characterised by low airspeeds. Using the doubly-labelled water technique, zebra finches Taeniopygia guttata engaging in repeated short flights were found to expend 13.65 kJ more than 'non-flying' controls, which equated to a flight expenditure of 27.8 times their basal metabolic rate. This is over three times the predicted flight expenditure derived from existing aerodynamic models. These data were used to determine a coefficient (0.11) for converting the mechanical power derived from aerodynamic models into metabolic power. An equation is presented, based on body mass, which can be used to predict the costs of short flights in ecological and behavioural studies of birds.


Association of short-term exposure to air pollution with depression in patients with sleep-related breathing disorders.
Air pollution is associated with sleep-related breathing disorders; however, the effects of air pollution on depression in patients with SRBDs remain unclear. A cross-sectional study was conducted to collect polysomnographic (PSG) data and Beck Depression Inventory-IA (BDI-IA) responses from 568 subjects with SRDBs in a sleep center in 2015 to 2017. Exposure to air pollution, including particulate matter with an aerodynamic diameter of ≤10 μm (PM10), particulate matter with an aerodynamic diameter of ≤2.5 μm (PM2.5), nitrogen (NO2), sulfur dioxide (SO2), carbon monoxide (CO) and ozone (O3), in 1-month averages was collected. Associations of air pollution with the respiratory disturbance index (RDI), oxygen desaturation index (ODI), arousal index (ARI), sleep architecture, and BDI-IA were examined. We observed that interquartile range (IQR) increases in 1-month PM2.5, PM10, and NO2 levels were respectively associated with 4.1/hour (h) (95% confidence interval (CI): 1.7/h to 6.4/h), 3.7/h (95% CI: 1.4/h to 6.0/h) and 1.9/h (95% CI: 0.1/h to 3.7/h) increases in the ARI. For sleep architecture, IQR increases in 1-month PM2.5 and CO levels were respectively associated with a 6.2% (95% CI: 6.1% to 6.3%) increase in non-rapid eye movement sleep 1 (N1) and a 2.0% (95% CI: -3.8% to -0.1%) decrease in non-rapid eye movement sleep 2 (N2). For depression, an IQR change in the 1-month CO was associated a moderate/severe depressive status according to the BDI-IA (odds ratio, OR: 2.981, p < 0.05; 95% CI: 1.032 to 8.611). Short-term exposure to air pollution increased the risk of arousal and light sleep as well as depression in patients with SRBDs. The results suggest that SRBD patients could be a population at risk for depression due to short-term exposure to air pollution.


In vivo detection of apoptotic cell death: a necessary measurement for evaluating therapy for myocarditis, ischemia, and heart failure.
If life is to continue, cells that have completed their useful function(s) must die in a timely manner. Apoptosis, programmed cell death, is a natural, orderly, energy-dependent process that causes cells to die without inducing an inflammatory response. In the heart, apoptosis plays pivotal roles in the development of myocarditis, cardiomyopathies, transplant rejection, the periinfarct zone in myocardial infarction, and reperfusion injury. Apoptosis is triggered either by a decrease in factors required to maintain the cell in good health or by an increase in factors which cause damage to the cell. When these factors tilt in the direction of death and the cell has sufficient time to respond, a common proteolytic cascade involving cysteine aspartic acid-specific proteases (caspases) is activated to initiate apoptosis. Cells that die by apoptosis autodigest their DNA and nuclear proteins, change the phospholipid composition on the outer surface of their cell membrane, and form lipid enclosed vesicles, which contain noxious intracellular contents, organelles, autodigested cytoplasm, and DNA. The compositional cell membrane phospholipid change that occurs with the onset of apoptosis is marked by the sudden expression of phosphatidylserine (PS), a phospholipid that ordinarily appears on the inner leaflet of the membrane, on the external leaflet of the membrane. The constant exposure of PS during apoptosis makes it an attractive target for radiopharmaceutical imaging. An endogenous human protein, annexin V, has a high affinity (kd = 7 nmol/L) for PS bound to the cell membrane. Fluorescence-labeled annexin V is used for histologic and cell-sorting studies to identify apoptotic cells. Annexin has been radiolabeled and binds to cells undergoing apoptosis in vivo. This review outlines some of the key features of apoptosis as contrasted to necrosis (unregulated cell death) and describes how these processes can be imaged with radionuclide techniques.


Hepatic microcirculation in the perfused cirrhotic rat liver.
Liver microcirculation in the perfused rat liver was assessed by the multiple indicator dilution technique. Comparative studies were carried out in noncirrhotic rats and in rats with cirrhosis secondary to chronic exposure to phenobarbital and carbon tetrachloride. The alterations of the sinusoidal bed were characterized by changes in the displacement of hepatic venous outflow curves of various diffusible substances (labeled albumin, sucrose, and water) relative to that of labeled erythrocytes (vascular reference). Outflow recoveries of lidocaine (a substance that penetrates the liver cell membrane freely and completely) and of labeled microspheres (15 microns diam) were also appraised. In all cirrhotic rats, unimodal erythrocytes and albumin curves were obtained. The sinusoidal space was significantly decreased when compared with normal rats (P less than 0.001) and the total space accessible to albumin became progressively restricted. In seven cirrhotic rats, the profiles of labeled sucrose and water curves were compatible with a flow-limited diffusion and the total distribution volumes were not significantly different from values found in noncirrhotic rats (P = NS), which indicates that sucrose and water were still able to diffuse into an extravascular space not accessible to albumin. In the other cirrhotic rats, labeled sucrose and water curves showed progressive bimodal changes not compatible with a flow-limited diffusion. Such alterations were not due to large intrahepatic shunts, since only 0.25% of the 15-microns microspheres were recovered in the outflow of cirrhotic rats. However, an early lidocaine outflow peak related in time to the peak erythrocyte curve was observed in cirrhotic, but not in noncirrhotic, rats. Lidocaine recovery varied greatly in cirrhotic rats and appeared to increase as the liver disease progressed. These data can be explained by capillarization of sinusoids and/or by the development of channels with poor permeability. Electron microscopic observations of these rat livers favored the latter. Thus, in cirrhotic rat liver, two kinds of alteration are likely: (a) the vascular space is decreased with collagenization of the extravascular space, limiting the diffusion of large molecules such as albumin; and (b) small channels with poorly permeable walls develop, limiting the diffusion of small molecules such as lidocaine, sucrose, and water. Large intrahepatic shunts are not a common feature.


Differentially expressed proteins in fluconazole-susceptible and fluconazole-resistant isolates of Candida glabrata.
The current study aimed to identify the differences presented in the proteome of fluconazole-susceptible isolates of Candida glabrata compared to those with fluconazole-resistant ones. Two-dimensional differential gel electrophoresis was applied to identify proteins that were differentially expressed in fluconazole-susceptible and fluconazole-resistant isolates of C. glabrata. Eight proteins including aspartyl-tRNA synthetase, translation elongation factor 3, 3-phosphoglycerate kinase, ribosomal protein L5, coproporphyrinogen III oxidase, pyruvate kinase, G-beta like protein, and F1F0-ATPase alpha subunit were found to be more abundantly represented, while four proteins including vitamin B12-(cobalamin)-independent isozyme of methionine synthase, microtubule-associated protein, adenylosuccinate synthetase, and aldose reductase were found to be less abundantly represented in fluconazole-resistant strains versus those with fluconazole-susceptible ones. These differentially expressed proteins were primarily associated with energy metabolism, stress response, and macromolecule synthesis. Proteins associated with energy metabolism, stress response, and macromolecule synthesis may play a role in the development of fluconazole resistance in the clinical isolates of C. glabrata. Multiple different mechanisms are involved in the development of fluconazole resistance in C. glabrata. These findings provide a scientific basis for discovering new genes and mechanisms associated with fluconazole resistance in C. glabrata.


[Characteristics of clinico-biochemical indicators in obese children with chronic tonsillitis].
Chronic tonsillitis (CT) was found in 63.1% of 190 obese children aged 3-15 years, 15% of them had no quinsy in the past. Onset of obesity was attributed to frequent quinsy in 18.3% of the examinees. Marked CT exacerbations were more typical for the children aged 6-7 years (10%). In 12-15-year-olds the disease tended to a sluggish course with scarce symptoms (25.8%). Pharyngoscopy discovered edema and hypertrophy of the tonsils in 74.1% of the patients. 35 patients underwent tonsillectomy. Examination of the removed tonsil tissue for phospholipids revealed pathological phospholipids lysophosphatidylcholine and phosphatidylinosite the quantities of which rose with growing obesity. It may be the result of negative action of obesity on tonsil structure.


3-deoxy-D-manno-2-octulosonic acid in the lipopolysaccharide of various strains of Pseudomonas cepacia.
Six clinical isolates of Pseudomonas cepacia (representing the five serotypes of the organism) were examined for the presence of 3-deoxy-D-manno-2-octulosonic acid (KDO) in their lipopolysaccharide (LPS). Purified LPS was examined for the presence of KDO by the thiobarbituric acid (TBA) assay and by gas chromatography. All strains possessed KDO. One strain possessed KDO that was detectable by the TBA assay after mild acid hydrolysis with 0.04 M H2SO4 at 100 degrees C for 20 min. The other strains also possessed KDO but it was only demonstrable by the TBA assay after strong acid hydrolysis (4 M HCl for 60 min at 100 degrees C). All six purified LPS preparations were shown to possess KDO by two separate gas chromatography procedures. LPS isolated from the six strains of P. cepacia was toxic for mice.


How I treat: the selection and acquisition of unrelated cord blood grafts.
Use of unrelated donor cord blood (CB) as an alternative stem cell source is increasing, and yet there is little information to guide transplant centers in the unique aspects of the search and selection of CB grafts. There is no mechanism to easily access the global inventory of CB units, nor is the product information provided by all banks standardized. To address these challenges, this manuscript reviews the logistics of the search, selection process, and acquisition of CB grafts as practiced by our center. Topics include who should be considered for a CB search, how to access the global CB inventory, and how to balance total nucleated cell dose and human leukocyte antigen match in unit selection. We discuss aspects of unit quality and other graft characteristics (processing methods, unit age, availability of attached segments, infectious disease, and hemoglobinopathy screening) to be considered. We incorporate these considerations into a unit selection algorithm, including how to select double-unit grafts. We also describe how we plan for unit shipment and the role of backup grafts. This review aims to provide a framework for CB unit selection and help transplantation centers perform efficient CB searches.


Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae): a review.
Lutzomyia longipalpis is the most important vector of AmericanVisceral Leishmaniasis (AVL) due to Leishmania chagasi in the New World. Despite its importance, AVL, a disease primarily of rural areas, has increased its prevalence and became urbanized in some large cities in Brazil and other countries in Latin America. Although the disease is treatable, other control measures include elimination of infected dogs and the use of insecticides to kill the sand flies. A better understanding of vector biology could also account as one more tool for AVL control. A wide variety of papers about L. longipalpis have been published in the recent past years. This review summarizes our current information of this particular sand fly regarding its importance, biology, morphology, pheromones genetics, saliva, gut physiology and parasite interactions.


Altered cortical CDC42 signaling pathways in schizophrenia: implications for dendritic spine deficits.
Spine density on the basilar dendrites of pyramidal neurons is lower in layer 3, but not in layers 5 and 6, in the dorsolateral prefrontal cortex (DLPFC) of subjects with schizophrenia. The expression of CDC42 (cell division cycle 42), a RhoGTPase that regulates the outgrowth of the actin cytoskeleton and promotes spine formation, is also lower in schizophrenia; however, CDC42 mRNA is lower across layers 3-6, suggesting that other lamina-specific molecular alterations are critical for the spine deficits in the illness. The CDC42 effector proteins 3 and 4 (CDC42EP3, CDC42EP4) are preferentially expressed in DLPFC layers 2 and 3, and CDC42EP3 appears to assemble septin filaments in spine necks. Therefore, alterations in CDC42EP3 could contribute to the lamina-specific spine deficits in schizophrenia. We measured transcript levels of CDC42, CDC42EP3, CDC42EP4; their interacting proteins (septins [SEPT2, 3, 5, 6, 7, 8, and 11], anillin), and other spine-specific proteins (spinophilin, PSD-95, and synaptopodin) in the DLPFC from 31 subjects with schizophrenia and matched normal comparison subjects. The expression of CDC42EP3 mRNA was significantly increased by 19.7%, and SEPT7 mRNA was significantly decreased by 6.9% in subjects with schizophrenia. Cortical levels of CDC42EP3 and SEPT7 mRNAs were not altered in monkeys chronically exposed to antipsychotic medications. Activated CDC42 is thought to disrupt septin filaments transiently in spine necks, allowing the molecular translocations required for synaptic potentiation. Thus, altered CDC42 signaling via CDC42EP3 may perturb synaptic plasticity and contribute to the spine deficits observed in layer 3 pyramidal neurons in schizophrenia.


Nostril sill augmentation in secondary cleft lip.
In most cases, after successful correction of primary cleft lip deformity, some problems remain to be solved. One of them is nostril sill depression, which should not be overlooked in cleft lip repair. However, there have been few reports on the anatomy of the depressed nostril sill and methods for augmenting the sill depression. The anatomy of the depressed nostril sill, its relationship to surrounding tissue, and the gross features of nostril sill depression in cleft lip deformity were reviewed, and a method in which a scar flap and a suture-fixation technique are used was developed for augmenting the depression. The clinical results of this method in 16 patients are presented.


A Bayesian meta-analysis of infants' ability to perceive audio-visual congruence for speech.
This paper quantifies the extent to which infants can perceive audio-visual congruence for speech information and assesses whether this ability changes with native language exposure over time. A hierarchical Bayesian robust regression model of 92 separate effect sizes extracted from 24 studies indicates a moderate effect size in a positive direction (0.35, CI [0.21: 0.50]). This result suggests that infants possess a robust ability to detect audio-visual congruence for speech. Moderator analyses, moreover, suggest that infants' audio-visual matching ability for speech emerges at an early point in the process of language acquisition and remains stable for both native and non-native speech throughout early development. A sensitivity analysis of the meta-analytic data, however, indicates that a moderate publication bias for significant results could shift the lower credible interval to include null effects. Based on these findings, we outline recommendations for new lines of enquiry and suggest ways to improve the replicability of results in future investigations.


Bioinspired mineralization of a functionalized injectable dense collagen hydrogel through silk sericin incorporation.
Collagen based hydrogels are frequently used as templates to mimic the native biomineralization process. However, a lack of structural control and their inherently poor mineralization capability represent challenges when used as bone-extracellular-matrix mimicking constructs. The aspiration-ejection of highly-hydrated collagen gels allows for their densification and fibrillar remodelling, leading to the production of injectable dense collagen (I-DC) gel scaffolds characterized by an osteoid-like structure. In this study, silk-extracted sericin (SS), a negatively-charged protein that is rich in anionic amino-acids such as Asp and Glu, was hybridized into I-DC gels to induce hydroxyapatite deposition and stimulate the osteoblastic differentiation of seeded mesenchymal stem cells (MSCs). The effect of SS content on the acellular mineralization of I-DC gels in simulated body fluid (SBF) and on modulating the proliferation and osteogenesis of seeded MSCs, in vitro, were investigated. Methylene blue staining indicated increasingly negatively charged gels through SS incorporation. Attributable to the carboxyl groups provided by the acidic SS amino-acids, serving as calcium-phosphate nucleation sites, there was a time dependent increase in hydroxyapatite deposition, approaching 90 wt% by day 14 in SBF. Three dimensionally seeded MSCs attached and proliferated in all gel types and SS-incorporation led to an increase in their metabolic activity. Relative to neat I-DC gels, alkaline phosphatase (at day 7), runt related transcription factor 2 (at day 21) and osteocalcin (at days 14 and 21) expression was higher in MSCs when seeded in SS-incorporated I-DC gels. Cell-induced mineralization was accelerated in SS-incorporated I-DC gels suggesting its osteostimulative potential. In sum, SS incorporation into clinically relevant I-DC gels can provide a strategy to design scaffolds with potential applications in bone tissue engineering.


Encorafenib, Binimetinib, and Cetuximab in BRAF V600E-Mutated Colorectal Cancer.
BRAFV600-mutated colorectal cancer (CRC) accounts for 8% to 12% of all CRC diagnoses. These tumors are often associated with specific patient features, including right-sided primary tumor location, peritoneal and non-regional lymph node involvement, and poor prognosis. In approximately 30% of cases, a simultaneous mismatch repair deficient (dMMR)/microsatellite instability-high (MSI-H) phenotype is identified. The prognostic impact of the BRAF mutation appears to be less marked in patients with MSI-H CRC than in patients with microsatellite stable (MSS) tumor. The treatment of BRAFV600-mutated CRC is still a challenge for the clinicians, mainly due to the poor survival outcomes obtained with traditional chemotherapy regimens. In recent years, two novel treatment strategies have offered remarkable changes in the treatment of this specific patient subgroup. The first approach has included targeted therapies directed against BRAF and MEK, with support from the epidermal growth factor receptor (EGFR) blockade. The second approach has included immunotherapeutic agents that have been shown to be particularly promising for patients with simultaneous dMMR/MSI-H phenotype. Here we review the clinical trials that specifically enrolled patients with BRAF-mutated CRC, from the phase I/II studies to the phase III trial BEACON CRC. We also examine the future directions towards a molecularly guided therapy for patients with BRAF-mutated CRC and the crucial role of a molecularly and clinically based algorithm in order to offer the best choice of treatment for these patients.


Predictive modeling of depression and anxiety using electronic health records and a novel machine learning approach with artificial intelligence.
Generalized anxiety disorder (GAD) and major depressive disorder (MDD) are highly prevalent and impairing problems, but frequently go undetected, leading to substantial treatment delays. Electronic health records (EHRs) collect a great deal of biometric markers and patient characteristics that could foster the detection of GAD and MDD in primary care settings. We approached the problem of predicting MDD and GAD using a novel machine learning pipeline to re-analyze data from an observational study. The pipeline constitutes an ensemble of algorithmically distinct machine learning methods, including deep learning. A sample of 4,184 undergraduate students completed the study, undergoing a general health screening and completing a psychiatric assessment for MDD and GAD. After explicitly excluding all psychiatric information, 59 biomedical and demographic features from the general health survey in addition to a set of engineered features were used for model training. We assessed the model's performance on a held-out test set and found an AUC of 0.73 (sensitivity: 0.66, specificity: 0.7) and 0.67 (sensitivity: 0.55, specificity: 0.7) for GAD, and MDD, respectively. Additionally, we used advanced techniques (SHAP values) to illuminate which features had the greatest impact on prediction for each disease. The top predictive features for MDD were being satisfied with living conditions and having public health insurance. The top predictive features for GAD were vaccinations being up to date and marijuana use. Our results indicate moderate predictive performance for the application of machine learning methods in detection of GAD and MDD based on EHR data. By identifying important predictors of GAD and MDD, these results may be used in future research to aid in the early detection of MDD and GAD.


Census-based mortality study of fertiliser manufactures.
This study was designed to investigate whether exposure to nitrate-containing dust during fertiliser manufacture was associated with an excess of deaths from cancer in general or specifically from cancers of the digestive tract, liver, lung, and bladder. It was based on data extracted from census schedules by the Office of Population Censuses and Surveys, occupational characteristics recorded by fertiliser workers at the 1961 and 1971 censuses of England and Wales being related to subsequent mortality ascertained through the National Health Service Central Register. The 1961 cohort, followed up until 1978, showed a "healthy worker effect" and no evidence of excess mortality from cancer at any site. The 1971 cohort also showed below average mortality during 1971-7 for all causes of death and for circulatory diseases, but there were more deaths from cancer than expected, due mainly to an excess of cancers of the lung and digestive tract. The excess of cancer was more pronounced, but not statistically significant, when compared with other employed men. Though the numbers for comparison were small, there was weak evidence of an association between cancer mortality and frequency of exposure to nitrate-containing dust in this cohort. It is difficult to reconcile the excess cancer mortality in the 1971 cohort with the more favorable level in the earlier cohort, since industrial hygiene has improved and the cohorts showed a similar distribution by region and social class. To examine further these conflicting results the 1971 cohort will be followed for a longer period and re-examined when more deaths have accrued.


A novel human homologue of Drosophila polycomblike gene is up-regulated in multiple cancers.
Polycomb group (PcG) proteins function to maintain the stable epigenetic repression of homeotic genes and other important developmental and cell cycle regulatory genes. Such maintenance establishes a form of cellular memory for its identity or state of differentiation. Accumulating evidence indicates that perturbation of this transcriptional memory may be required for tumor progression and may represent a hallmark of cancer. We have identified a novel gene encoding a human homologue of the Drosophila polycomblike protein, hPCL3. Through alternative polyadenylation and/or splicing, the gene encodes two nuclear proteins, hPCL3S and hPCL3L. Both proteins repressed transcription upon recruitment to the proximity of an HSV-tk promoter by a Gal4 DNA binding domain. Interestingly, the products of the hPCL3 gene, particularly the short form, hPCL3S, are markedly overexpressed in many types of cancers, including colon, skin, lung, rectal, cervical, uterus, and liver cancers. This increase in expression correlated with tumor progression. Both hPCL3S and hPCL3L messages were increased dramatically in most cell lines derived from various stages of melanoma and glioma tumor progression. Thus, our data link PcG deregulation to the progression of multiple cancers and may have important implications for unraveling the mechanisms of tumor progression.


The functional anatomy of divided attention in amnestic mild cognitive impairment.
Recent neuroimaging studies have demonstrated changes in brain function in cognitively normal subjects at increased risk of developing Alzheimer's disease. Amnestic mild cognitive impairment (AMCI) carries a high risk of developing into Alzheimer's disease. In AMCI altered cortical activation has been demonstrated during memory tasks, using functional MRI (fMRI). Memory and attention are closely related cognitive functions. It is unclear whether the memory impairment of AMCI is associated with attentional deficits of the sort likely to be revealed by tasks requiring divided attention. Ten older adults (mean age 72 years, range 57-81 years) with AMCI were compared with healthy matched controls on divided attention and passive sensory processing tasks using fMRI. During the divided attention task both groups activated similar regions of left hemispheric prefrontal and extrastriate visual cortex. However, the AMCI group had attenuated prefrontal activation compared with age matched controls. On the passive sensory processing task there was no difference between the AMCI and control groups. We conclude that there are changes in the functional network subserving divided attention in patients with AMCI as reflected in the attenuation of prefrontal cortical activation. These findings have implications for evaluating cognition in AMCI and also for monitoring the effects of future treatments in AMCI.


Tablet-Based eBooks for Young Children: What Does the Research Say?
Young children's use of electronic books (eBooks) is increasing as handheld touch screen devices, such as tablets, become increasingly available. Although older children's reading on tablets has been more broadly investigated, less is known about the impacts of digital reading for infant, toddlers, and preschoolers. This review compares the educational affordances of reading on tablets versus print books for young children's learning. A qualitative synthesis of research on tablet-based eReading and young children's learning from screens was conducted. When eBooks are designed well, preschool-aged children learn equally well and sometimes more than from print books. However, enhanced eBooks with sounds, animations, and games can distract children and reduce learning. When book-sharing with an adult, conversations during eBook reading are often about the platform while print book conversations are more often about the book content. For young children (0-2 yr), there is a paucity of research, but broader studies on learning from screens suggest limited educational benefits of tablet use for this age group. The authors recommend that (1) the selection of eBooks (especially enhanced eBooks) be thoughtful as games or animations that are not related to the story content can be distracting for young children, (2) adults share in the reading experience as discussions of the story, text, and characters have been found to enhance comprehension, language development, and print awareness, and (3) tablet eBook use be restricted for infants and toddlers, as they benefit more from face-to-face interaction with caregivers than from interactive screens alone.


isiKnock: in silico knockouts in signaling pathways.
isiKnock is a new software that automatically conducts in silico knockouts for mathematical models of signaling pathways. The software allows for the prediction of the behavior of biological systems after single or multiple knockout. The implemented algorithm applies transition invariants and the novel concept of Manatee invariants. A knockout matrix visualizes the results. The tool enables the analysis of dependencies, for example, in signal flows from the receptor activation to the cell response at steady state. isiKnock is an open-source tool, freely available at http://www.bioinformatik.uni-frankfurt.de/tools/isiKnock/. It requires at least Java 8 and runs under Microsoft Windows, Linux, and Mac OS.


Depletion of Caveolin-1 in Type 2 Diabetes Model Induces Alzheimer's Disease Pathology Precursors.
Type 2 diabetes mellitus (T2DM) is a risk factor for the development of late-onset Alzheimer's disease (AD). However, the mechanism underlying the development of late-onset AD is largely unknown. Here we show that levels of the endothelial-enriched protein caveolin-1 (Cav-1) are reduced in the brains of T2DM patients compared with healthy aging, and inversely correlated with levels of β-amyloid (Aβ). Depletion of Cav-1 is recapitulated in the brains of db/db (Leprdb ) diabetic mice and corresponds with recognition memory deficits as well as the upregulation of amyloid precursor protein (APP), BACE-1, a trending increase in β-amyloid Aβ42/40 ratio and hyperphosphorylated tau (p-tau) species. Importantly, we show that restoration of Cav-1 levels in the brains of male db/db mice using adenovirus overexpressing Cav-1 (AAV-Cav-1) rescues learning and memory deficits and reduces pathology (i.e., APP, BACE-1 and p-tau levels). Knocking down Cav-1 using shRNA in HEK cells expressing the familial AD-linked APPswe mutant variant upregulates APP, APP carboxyl terminal fragments, and Aβ levels. In turn, rescue of Cav-1 levels restores APP metabolism. Together, these results suggest that Cav-1 regulates APP metabolism, and that depletion of Cav-1 in T2DM promotes the amyloidogenic processing of APP and hyperphosphorylation of tau. This may suggest that depletion of Cav-1 in T2DM underlies, at least in part, the development of AD and imply that restoration of Cav-1 may be a therapeutic target for diabetic-associated sporadic AD.SIGNIFICANCE STATEMENT More than 95% of the Alzheimer's patients have the sporadic late-onset form (LOAD). The cause for late-onset Alzheimer's disease is unknown. Patients with Type 2 diabetes mellitus have considerably higher incidence of cognitive decline and AD compared with the general population, suggesting a common mechanism. Here we show that the expression of caveolin-1 (Cav-1) is reduced in the brain in Type 2 diabetes mellitus. In turn, reduced Cav-1 levels induce AD-associated neuropathology and learning and memory deficits. Restoration of Cav-1 levels rescues these deficits. This study unravels signals underlying LOAD and suggests that restoration of Cav-1 may be an effective therapeutic target.


Stereotactic Ablative Radiotherapy (SABR) in inoperable oligometastatic disease from colorectal cancer: a safe and effective approach.
To assess the safety and efficacy of Stereotactic Ablative Radiotherapy (SABR) in oligometastatic patients from colorectal cancer. 82 patients with 1-3 inoperable metastases confined to one organ (liver or lung), were treated with SABR for a total of 112 lesions in an observational study. Prescription dose ranged between 48 and 75 Gy in 3 or 4 consecutive fractions. Primary end-points were local control (LC), overall survival (OS) and progression-free survival (PFS). Secondary end-point was toxicity. Median follow-up was 24 months (range 3-47). One, two and three years LC rate was 90%,80% and 75% (85%,75% and 70% for lung and 95%, 90% and 85% for liver metastases; no statistically significance was found). The difference in LC between the subgroup of lesions treated with ≥ 60 Gy (n = 58) and those irradiated with <60 Gy (n = 52) was statistically significant, with a 1, 2 and 3 yrs LC of 97%,92% and 83% for the higher dose, compared to 85%,70% and 70% for the lower dose (p < 0.04). Median OS was 32 months. Actuarial OS rate at 1, 2 and 3 yrs was 85%,65% and 43%. Univariate analysis showed a correlation only between OS and cumulative GTV > 3 cm (p < 0.02). Median PFS was 14 months, with a PFS rate of 56% at 1 yr and 40% at 2-3 yrs, without correlation with the site and prescription dose (p < 0.48 and p < 0.56). No patients experienced radiation-induced liver disease or grade >3 toxicity. SABR is a safe and feasible alternative treatment of oligometastatic colorectal liver and lung metastases in patients not amenable to surgery or other ablative treatments.


European stem cell research in legal shackles.
Advances in stem cell biology have raised legal challenges to the patentability of stem cells and any derived technologies and processes. In 1999, Oliver Brüstle was granted a patent for the generation and therapeutic use of neural cells derived from human embryonic stem cells (hESCs). The patent was challenged and put before the European Court of Justice, which ruled that inventions involving the prior destruction of human embryos cannot be patented. The legal maneuvering around this case demonstrates that the future of stem cell-based patents in Europe remains unsettled. Furthermore, owing to the European Court's broad definition of hESC as 'any cell that is capable of commencing development into a human being,' novel technologies that could eliminate the need for hESCs, such as induced pluripotent stem cells (iPSCs), are at risk of being included under the same ruling. Advances in the in vitro development of germ cells from pluripotent stem cells may one day provide a direct developmental path from iPSC to oocyte and sperm, and, according to the European Court's reasoning, legally equate iPSCs with human embryos. In this review, we will briefly discuss the Brüstle v Greenpeace case and the implications of the European Court of Justice's ruling. We will identify potential risks for stem cell research and future therapeutics resulting from the broad legal definition of the human embryo. Finally, we will broach the current legal landscape, as this broad definition has also created great uncertainty about the status of human iPSCs.


A two-stage reconstruction of fluorescence molecular tomography based on sparse regularization.
Fluorescence molecular tomography (FMT) is a promising imaging modality that offers the possibilities to monitor cellular and molecular function in vivo. However, accurate and stable reconstruction of fluorescence-labeled targets remains a challenging problem. In this contribution, a two-stage reconstruction algorithm that combines sparse regularization with adaptive finite element method is proposed, and two different inversion algorithms are employed separately on the initial coarse mesh and the second refined one. Numerical experiment results with a digital mouse model demonstrate the stability and computational efficiency of the proposed method for FMT.


Free fatty acid receptor 4 inhibitory signaling in delta cells regulates islet hormone secretion in mice.
Maintenance of glucose homeostasis requires the precise regulation of hormone secretion from the endocrine pancreas. Free fatty acid receptor 4 (FFAR4/GPR120) is a G protein-coupled receptor whose activation in islets of Langerhans promotes insulin and glucagon secretion and inhibits somatostatin secretion. However, the contribution of individual islet cell types (α, β, and δ cells) to the insulinotropic and glucagonotropic effects of GPR120 remains unclear. As gpr120 mRNA is enriched in somatostatin-secreting δ cells, we hypothesized that GPR120 activation stimulates insulin and glucagon secretion via inhibition of somatostatin release. Glucose tolerance tests were performed in mice after administration of selective GPR120 agonist Compound A. Insulin, glucagon, and somatostatin secretion were measured in static incubations of isolated mouse islets in response to endogenous (ω-3 polyunsaturated fatty acids) and/or pharmacological (Compound A and AZ-13581837) GPR120 agonists. The effect of Compound A on hormone secretion was tested further in islets isolated from mice with global or somatostatin cell-specific knock-out of gpr120. Gpr120 expression was assessed in pancreatic sections by RNA in situ hybridization. Cyclic AMP (cAMP) and calcium dynamics in response to pharmacological GPR120 agonists were measured specifically in α, β, and δ cells in intact islets using cAMPER and GCaMP6 reporter mice, respectively. Acute exposure to Compound A increased glucose tolerance, circulating insulin, and glucagon levels in vivo. Endogenous and/or pharmacological GPR120 agonists reduced somatostatin secretion in isolated islets and concomitantly demonstrated dose-dependent potentiation of glucose-stimulated insulin secretion and arginine-stimulated glucagon secretion. Gpr120 was enriched in δ cells. Pharmacological GPR120 agonists reduced cAMP and calcium levels in δ cells but increased these signals in α and β cells. Compound A-mediated inhibition of somatostatin secretion was insensitive to pertussis toxin. The effect of Compound A on hormone secretion was completely absent in islets from mice with either global or somatostatin cell-specific deletion of gpr120 and partially reduced upon blockade of somatostatin receptor signaling by cyclosomatostatin. Inhibitory GPR120 signaling in δ cells contributes to both insulin and glucagon secretion in part by mitigating somatostatin release.


CO chemisorption and dissociation at high coverages during CO hydrogenation on Ru catalysts.
Density functional theory (DFT) and infrared spectroscopy results are combined with mechanism-based rate equations to assess the structure and thermodynamics of chemisorbed CO (CO*) and its activation during Fischer-Tropsch synthesis (FTS). CO* binding becomes weaker with increasing coverage on Ru(0001) and Ru201 clusters, but such decreases in binding energy occur at higher coverages on Ru201 clusters than on Ru(0001) surfaces (CO*/Ru = 1.55 to 0.75); such differences appear to reflect weaker repulsive interactions on the curved surfaces prevalent on small Ru201 clusters. Ru201 clusters achieve stable supramonolayer coverages (CO*/Ru > 1) by forming geminal dicarbonyls at low-coordination corner/edge atoms. CO* infrared spectra on Ru/SiO2 (~7 nm diameter) detect mobile adlayers that anneal into denser structures at saturation. Mechanism-based FTS rate equations give activation energies that reflect the CO*-saturated surfaces prevalent during catalysis. DFT-derived barriers show that CO* predominantly reacts at (111) terraces via H-assisted reactions, consistent with measured effects of H2 and CO pressures and cluster size effects on rates and O-rejection selectivities. Barriers are much higher for unassisted CO* dissociation on (111) terraces and low-coordination atoms, including step-edge sites previously proposed as active sites for CO* dissociation during FTS. DFT-derived barriers indicate that unassisted CO* dissociation is irreversible, making such steps inconsistent with measured rates. The modest activation barriers of H-assisted CO* dissociation paths remove a requirement for special low-coordination sites for unassisted CO* activation, which is inconsistent with higher rates on larger clusters. These conclusions seem generally applicable to Co, Fe, and Ru catalysts, which show similar FTS rate equations and cluster size effects. This study also demonstrates the feasibility and relevance of DFT treatments on the curved and crowded cluster surfaces where catalysis occurs.


Blood Conservation Using Tranexamic Acid Is Not Superior to Epsilon-Aminocaproic Acid After Total Knee Arthroplasty.
Epsilon-aminocaproic acid (EACA) and tranexamic acid (TXA) are synthetic amino acid derivatives that interfere with fibrinolysis, promoting hemostasis by pharmacological means. Although both drugs have been shown to decrease blood loss with a minimal risk of thromboembolic adverse events following cardiac and vascular surgery, we are aware of only 1 published trial that directly compared the antifibrinolytic effects of EACA with those of TXA after total knee arthroplasty (TKA). The primary aim of this prospective, randomized, controlled trial was to determine whether TXA provides superior blood conservation following TKA compared with that provided by EACA. A total of 194 patients scheduled to undergo a primary unilateral TKA in the same community-based hospital were prospectively randomized to receive intravenous EACA (n = 96) or TXA (n = 98). Both the patients and the operating surgeons were blinded to the treatment assignments. Primary outcome measures included transfusions, estimated blood loss, and the drop in the hemoglobin (Hgb) level. Secondary outcomes measures included the change in the serum creatinine level, postoperative complications, and length of hospital stay. Although the patients who received TXA averaged less estimated blood loss than the patients who received EACA (t185 = 2.18, p = 0.031; mean difference = 144.2 mL, 95% confidence interval = 13.62 to 274.78 mL), no transfusions were required in either group. We observed no statistically significant or clinically relevant between-group differences in the change in Hgb or serum creatinine level, postoperative complications, or length of hospital stay. Although the estimated blood loss was significantly greater in the EACA group, no transfusions were required and no significant between-group differences were observed for any other outcomes measured. We concluded that EACA may be an acceptable alternative to TXA for blood conservation following TKA, although replication of our results in noninferiority trials is necessary. Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.


High blood pressure. detection and treatment by general practitioners.
A questionnaire was sent to a 10% random sample of general practitioners in England and Wales on their attitudes to the detection and treatment of hypertension; 62% responded and no further inquiry was made. Their view on detection and criteria for treatment and investigations performed were considered in relation to their background. More of the older practitioners always measured blood pressure and 36% of all practitioners believed that hypertensive patients usually present with symptoms. Altogether 91% thought that strokes could be prevented by treating hypertension, and only 18% reported difficulty in keeping patients on treatment. Older practitioners preferred to measure the distolic pressure using phase five, while the younger preferred phase four. Nearly all doctors were satisfied with their current sphygmomanometers.


Interleukin-1 alpha modulates luteinizing hormone stimulated cyclic AMP and progesterone release from human granulosa cells in vitro.
This study examines the possible direct effect of interleukin-1 alpha (IL-1 alpha) upon human granulosa cells. The cells were isolated from follicles of stimulated cycles in women undergoing oocyte retrieval for in-vitro fertilization. Purified cell preparations were cultured for different time periods in the presence of IL-1 alpha and human luteinizing hormone (LH) or follicle stimulating hormone. IL-1 alpha stimulated basal as well as LH-induced progesterone accumulation. The response in terms of cyclic AMP was more complex, there was no effect of IL-1 alpha on basal cyclic AMP accumulation. However, at the highest concentration tested (50 IU/ml), IL-1 alpha enhanced cyclic AMP accumulation over that seen with LH alone. At a lower concentration, IL-1 alpha either had no effect or was slightly inhibitory to the LH-induced cyclic AMP accumulation, depending on the culture period. Our results, taken together with other findings, are compatible with the view that IL-1 alpha has a potential regulatory role in the granulosa-luteal cell transition in the human ovary.


Bond Polarizability Model for Sum Frequency Generation at the Al2O3(0001)-H2O Interface.
Sum Frequency Generation (SFG) is a powerful, surface-specific vibrational probe ideally suited to studying buried interfaces; however, insight from theory is often necessary to explain the microscopic origins of the spectral features. To calculate the SFG spectrum at an insulating solid/liquid interface, we develop a flexible polarizability model that takes local dipole interactions into account, rather than assuming additive polarizabilities. We use this model to calculate bond dipoles and polarizabilities that reflect the local geometry of the interface. We apply our method to the Al2O3(0001)-H2O interface, where we reproduce the experimental spectrum and show the two H stretching peaks come from solvent and surface modes separately, not from H2O molecules with different coordination numbers as previously thought. Our work therefore emphasizes the importance of treating both surface and solvent at the same level of theory for accurate spectroscopic calculations.


Controlling chemical self-assembly by solvent-dependent dynamics.
The influence of the ratio between poor and good solvent on the stability and dynamics of supramolecular polymers is studied via a combination of experiments and simulations. Step-wise addition of good solvent to supramolecular polymers assembled via a cooperative (nucleated) growth mechanism results in complete disassembly at a critical good/poor solvent ratio. In contrast, gradual disassembly profiles upon addition of good solvent are observed for isodesmic (non-nucleated) systems. Due to the weak association of good solvent molecules to monomers, the solvent-dependent aggregate stability can be described by a linear free-energy relationship. With respect to dynamics, the depolymerization of π-conjugated oligo(p-phenylene vinylene) (OPV) assemblies in methylcyclohexane (MCH) upon addition of chloroform as a good solvent is shown to proceed with a minimum rate around a critical chloroform/MCH solvent ratio. This minimum disassembly rate bears an intriguing resemblance to phenomena observed in protein unfolding, where minimum rates are observed at the thermodynamic midpoint of a protein denaturation experiment. A kinetic nucleation-elongation model in which the rate constants explicitly depend on the good solvent fraction is developed to rationalize the kinetic traces and further extend the insights by simulation. It is shown that cooperativity, i.e., the nucleation of new aggregates, plays a key role in the minimum polymerization and depolymerization rate at the critical solvent composition. Importantly, this shows that the mixing protocol by which one-dimensional aggregates are prepared via solution-based processing using good/poor solvent mixtures is of major influence on self-assembly dynamics.


Disruption of the Bcchs3a chitin synthase gene in Botrytis cinerea is responsible for altered adhesion and overstimulation of host plant immunity.
The fungal cell wall is a dynamic structure that protects the cell from different environmental stresses suggesting that wall synthesizing enzymes are of great importance for fungal virulence. Previously, we reported the isolation and characterization of a mutant in class III chitin synthase, Bcchs3a, in the phytopathogenic fungus Botrytis cinerea. We demonstrated that virulence of this mutant is severely impaired. Here, we describe the virulence phenotype of the cell-wall mutant Bcchs3a on the model plant Arabidopsis thaliana and analyze its virulence properties, using a variety of A. thaliana mutants. We found that mutant Bcchs3a is virulent on pad2 and pad3 mutant leaves defective in camalexin. Mutant Bcchs3a was not more susceptible towards camalexin than the wild-type strain but induced phytoalexin accumulation at the infection site on Col-0 plants. Moreover, this increase in camalexin was correlated with overexpression of the PAD3 gene observed as early as 18 h postinoculation. The infection process of the mutant mycelium was always delayed by 48 h, even on pad3 plants, probably because of lack of mycelium adhesion. No loss in virulence was found when Bcchs3a conidia were used as the inoculum source. Collectively, these data led us to assign a critical role to the BcCHS3a chitin synthase isoform, both in fungal virulence and plant defense response.


The nuclease sensitivity of active genes.
Brief micrococcal nuclease digestion of chick embryonic red blood cells results in preferential excision and solubilization of monomer nucleosomes associated with beta-globin sequences and also 5'-sequences flanking the beta-globin gene. Both regions are DNAse-I sensitive in nuclei. Such salt-soluble nucleosomes are enriched in all four major HMG proteins but HMG1 and 2 are only weakly associated. These nucleosomes appear to have lost much of the DNAse-I sensitivity of active genes. The HMG14 and 17-containing salt-soluble nucleosomes separated by electrophoresis are not DNAse-I sensitive and contain inactive gene sequences as well as active sequences. Reconstitution of HMG proteins onto bulk nucleosomes or chromatin failed to reveal an HMG-dependent sensitivity of active genes as assayed by dot-blot hybridization and it was found that the DNAse-I sensitivity of ASV proviral sequences as assayed by dot-blot hybridization was not HMG-dependent. These results indicate that higher order chromatin structures might be responsible for nuclease sensitivity of active genes.


Immunolocalization of Ki-67 in different periodontal conditions.
Ki-67 which is a non-histone nuclear protein which is expressed in proliferating cells, during all the active phases of the cell cycle. Increased Ki-67 expression has been seen in several inflammatory and malignant conditions like diabetes, rheumatoid arthritis, atherosclerosis, pancreatitis and squamous cell carcinoma. The aim of the present study is to analyze the expression of Ki-67 in gingival tissues by immunohistochemistry in smokers and non-smokers with healthy gingiva and chronic periodontitis. Gingival biopsies (n = 32) were obtained from smokers who had clinically healthy gingiva (n = 8), smokers with periodontitis (n = 8), chronic periodontitis (n = 8) and healthy gingiva (n = 8). The expression of Ki-67 was evaluated immunohistochemically. Mean and standard deviation were estimated for the gingival tissue extract sample for each study group. Mean values were compared between different study groups by, one way ANOVA, post hoc analysis. In this study P < 0.05 was considered as the level of significance. The mean number of Ki-67 positive cells/field was higher in the smokers with periodontitis group. When the mean Ki-67 positive cells were compared between different groups, statistical significant difference was observed between healthy and both the periodontitis groups (P = 0.000) and between smokers group (P = 0.001). Ki-67 was maximally expressed in smoker with periodontitis followed by chronic periodontitis patients, healthy smokers and healthy control patients which shed light on the toxic effects of tobacco in dysregulating the cell cycle and cellular proliferation. The findings of this study also help us to understand the role of the cell cycle in resolution of periodontal inflammation which is a salient feature in the pathogenesis of chronic periodontitis.


Cellular immune mechanisms in inflammatory myopathies.
The inflammatory myopathies include dermatomyositis, polymyositis, and inclusion body myositis. In dermatomyositis, muscle fiber injury is secondary to an antibody- or immune-complex-mediated immune response against a vascular-endothelial component. In polymyositis and inclusion body myositis, CD8+ T cells and macrophages invade and eventually destroy initially nonnecrotic muscle fibers. The autoaggressive T cells have the phenotype of activated (HLA-DR+) memory (CD45RO+) cells. T-cell receptor analyses indicate that the autoaggressive T cells are oligoclonal. In inflammatory lesions, muscle fibers express various cytoplasmic and surface molecules that are not detectable in normal fibers. These molecules, which include HLA class I antigens, heat-shock proteins, adhesion molecules, and Fas, are probably induced by locally secreted cytokines. The autoaggressive CD8+ T cells harbor granules containing perforin that aggregate near the contact zone with the target muscle fiber. This is consistent with a perforin- and secretion-dependent mechanism of muscle fiber injury. Many invaded muscle fibers also express the Fas "death receptor," but signs of apoptosis are absent.


Decoding the enigma: unveiling the molecular transmission of avian-associated tet(X4)-positive E. coli in Sichuan Province, China.
Tigecycline is considered one of the "last resort antibiotics" for treating complex infections caused by multidrug-resistant (MDR) bacteria, especially for combating clinical resistant strains that produce carbapenemases. However, the tet(X4) gene, which carried by different plasmids can mediate high levels of bacterial resistance to tigecycline, was first reported in 2019. Here, we report the emergence of the plasmid-mediated tet(X4) in avian environment of Sichuan Province. A total of 21 tet(X4)-positive Escherichia coli (E. coli) strains were isolated and identified from avian samples in selected regions, with an isolation rate of 1.6% (21/1,286), and all of them were MDR strains. Multilocus Sequence Typing (MLST) method was used to classify the 21 tet(X4)-positive E. coli into the ST206, ST761, ST155, ST1638, ST542, and ST767 types, which also belong to the 3 phylogenetic subgroups A, B1, and C. Tet(X4) is located on mobile plasmids that can be efficiently and stably propagated. The results of fitness cost experiments showed that tet(X4)-positive plasmids may incur some fitness cost to host bacteria, but different tet(X4)-positive plasmids bring about differential fitness costs. Whole-genome sequencing further confirmed the tet(X4) gene can be located on IncX1-type plasmids and the core genetic structures are ISVsa3-rdmc-tet(X4) or rdmc-tet(X4)-ISVsa3, the former is a 7 copies tandem repeat structure. In this study, we isolated and identified tet(X4)-positive E. coli from the avian origin in Sichuan, analyzed the mobility of the tet(X4) by conjugational transfer and S1-PFGE, and evaluated the biological characteristics of the tet(X4)-positive plasmid using the results of conjugational frequency, plasmid stability, and fitness costs. Finally, combined with the third-generation whole-genome sequencing analysis, the molecular transmission characteristics of the tet(X4) were preliminarily clarified, providing a scientific basis for guiding veterinary clinical use in this area, as well as risk assessment and prevention of the transfer and spread of tigecycline resistant strains or genes.


Toxic accumulation of alpha-ketobutyrate caused by inhibition of the branched-chain amino acid biosynthetic enzyme acetolactate synthase in Salmonella typhimurium.
Biochemical and genetic analyses of the bacterium Salmonella typhimurium suggest that accumulation of alpha-ketobutyrate partially mediates the herbicidal activity of acetolactate synthase inhibitors. Growth inhibition of wild-type bacteria by the herbicide sulfometuron methyl was prevented by supplementing the medium with isoleucine, an allosteric inhibitor of threonine deaminase-catalyzed synthesis of alpha-ketobutyrate. In contrast, isoleucine did not rescue the growth of a mutant containing a threonine deaminase unresponsive to isoleucine. Moreover, the hypersensitivity of seven Tn10 insertion mutants to growth inhibition by sulfometuron methyl and alpha-ketobutyrate correlated with their inability to convert alpha-ketobutyrate to less noxious metabolites. We propose that alpha-ketobutyrate accumulation is an important component of sulfonylurea and imidazolinone herbicide action.


Human mitochondrial RNA polymerase: evaluation of the single-nucleotide-addition cycle on synthetic RNA/DNA scaffolds.
The human mitochondrial RNA polymerase (h-mtRNAP) serves as both the transcriptase for expression and the primase for replication of mitochondrial DNA. As such, the enzyme is of fundamental importance to cellular energy metabolism, and defects in its function may be related to human disease states. Here we describe in vitro analysis of the h-mtRNAP kinetic mechanism for single, correct nucleotide incorporation. This was made possible by the development of efficient methods for expression and purification of h-mtRNAP using a bacterial system and by utilization of assays that rely on simple, synthetic RNA/DNA scaffolds without the need for mitochondrial transcription accessory proteins. We find that h-mtRNAP accomplishes single-nucleotide incorporation by using the same core steps, including conformational change steps before and after chemistry, that are prototypical for most types of nucleic acid polymerases. The polymerase binds to scaffolds via a two-step mechanism consisting of a fast initial-encounter step followed by a much slower isomerization that leads to catalytic competence. A substantial solvent deuterium kinetic isotope effect was observed for the forward reaction, but none was detectable for the reverse reaction, suggesting that chemistry is at least partially rate-limiting in the forward direction but not in the reverse. h-mtRNAP appears to exercise much more stringent surveillance over base than over sugar in determining the correctness of a nucleotide. The utility of developing the robust in vitro assays described here and of establishing a baseline of kinetic performance for the wild-type enzyme is that biological questions concerning h-mtRNAP may now begin to be addressed.


Anaerobic sulfur metabolism coupled to dissimilatory iron reduction in the extremophile Acidithiobacillus ferrooxidans.
Gene transcription (microarrays) and protein levels (proteomics) were compared in cultures of the acidophilic chemolithotroph Acidithiobacillus ferrooxidans grown on elemental sulfur as the electron donor under aerobic and anaerobic conditions, using either molecular oxygen or ferric iron as the electron acceptor, respectively. No evidence supporting the role of either tetrathionate hydrolase or arsenic reductase in mediating the transfer of electrons to ferric iron (as suggested by previous studies) was obtained. In addition, no novel ferric iron reductase was identified. However, data suggested that sulfur was disproportionated under anaerobic conditions, forming hydrogen sulfide via sulfur reductase and sulfate via heterodisulfide reductase and ATP sulfurylase. Supporting physiological evidence for H2S production came from the observation that soluble Cu(2+) included in anaerobically incubated cultures was precipitated (seemingly as CuS). Since H(2)S reduces ferric iron to ferrous in acidic medium, its production under anaerobic conditions indicates that anaerobic iron reduction is mediated, at least in part, by an indirect mechanism. Evidence was obtained for an alternative model implicating the transfer of electrons from S(0) to Fe(3+) via a respiratory chain that includes a bc(1) complex and a cytochrome c. Central carbon pathways were upregulated under aerobic conditions, correlating with higher growth rates, while many Calvin-Benson-Bassham cycle components were upregulated during anaerobic growth, probably as a result of more limited access to carbon dioxide. These results are important for understanding the role of A. ferrooxidans in environmental biogeochemical metal cycling and in industrial bioleaching operations.


Wearability and physiological effects of custom-fitted vs self-adapted mouthguards.
The purpose of this study was to measure the comfort, wearability, physiological effects and its influence on athletes' physical performance, of custom-fitted compared with self-adapted mouthguards (MGs). Eleven rugby players were put under specific efforts similar to those of the competition. Each player made three consecutive tests randomly wearing a commercially available 'boil-and-bite' self-adapted mouthguard (MG2), a custom-fitted mouthguard (MG3), and no mouthguard (reference). Forced expiratory air volume at 1 s (FEV(1)), expiratory flow rates peak (PEF), forced vital capacity (FVC), rebound (RB) jump 15 s, and counter-movement jump (CMJ) were measured on each player before and after the training exercise tests. Subjective evaluations by means of a visual analog scale (VAS) questionnaire took place. Comforts, adaptability, stability, tiredness, thirst, oral dryness, nausea, ability to talk, breathe, and drink were evaluated. The wearing of the self-adapted MG showed significant improvement in PEF (P < 0.05). There were no statistically significance differences regarding the others spirometer parameters. In CMJ, there were no differences between both the MGs. On RB power was similar with both MGs and control. However, RB height reduced significantly wearing MGs. MG3 showed superior properties in comfort, adaptability, stability, and ability to talk and to breathe. MG3 showed the smallest range of changes in players' performance, suggesting improved fit, comfort, and acceptation compared with MG2. Furthermore, its greatest advantage is the individualized design according to the proper anatomy of the oral cavity. Greater efforts must be made to improve the comfort of MGs if their use is to be increased.


Analysis of heat loss mechanisms for mobile tent-type refuge alternatives.
Federal regulations require that refuge alternatives (RAs) be located within 305 m (1,000 ft) of the working face and spaced at one-hour travel distances in the outby area in underground coal mines, in the event that miners cannot escape during a disaster. The Mine Safety and Health Administration mandates that RAs provide safe shelter and livable conditions for a minimum of 96 hours while maintaining the apparent temperature below 35 °C (95 °F). The U.S. National Institute for Occupational Safety and Health used a validated thermal simulation model to examine the mechanisms of heat loss from an RA to the ambient mine and the effect of mine strata composition on the final internal dry bulb temperature (DBT) for a mobile tent-type RA. The results of these studies show that 51 percent of the heat loss from the RA to the ambient mine is due to radiation and 31 percent to conduction. Three mine width and height configurations and four mine strata compositions were examined. The final DBT inside the RA after 96 hours varied by less than 1 °C (1.8 °F) for the three mine width/height configurations and by less than 2 °C (3.6 °F) for the four mine strata compositions.


Immunophenotypings of malignant epithelial mesothelioma and their roles in the differential diagnosis.
To investigate the immunophenotypings of malignant epithelial mesothelioma (MEM), and to seek the valuable markers in distinguishing peritoneal MEM from peritoneal metastatic ovarian adenocarcinoma (OA) and colorectal adenocarcinoma (CA), immunohistochemical SP method was used to detect expressions of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 in paraffin-embedded tissues of 18 cases of MEM, 20 OA and 20 CA. The results showed that there was a significant difference in the expressions of E-cadherin, CA19-9 and MOC-31 between MEM and OA group (P<0.05). Similarly, the difference in the expression of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 between MEM and CA groups is significant (P<0.05). These results indicate that HBME-1 could be used as a positive marker in distinguishing MEM from CA. E-cadherin, CA19-9 and MOC-31 are considered to be useful negative markers in diagnostic distinction between MEM and metastatic adenocarcinomas, including OA and CA. CK7 is the best positive marker in distinguishing MEM from CA, but this marker appears to be valueless in discriminating MEM from OA.


Accreditation: a tool for organizational change in hospitals?
Examines the dynamics of change that operated following preparations for accreditation. The study was conducted from May 1995 to October 2001 in a university hospital center in France after the introduction in 1996 of mandatory accreditation. An embedded explanatory case study sought to explore the organizational changes: a theoretical framework for analyzing change was developed; semi-structured interviews, focus groups, and questionnaires addressed to the hospital's professionals were used and documents were collected; and qualitative and quantitative analyses were carried out. Professionals from clinical and medico-technical departments participated most. Preparations for accreditation provided an opportunity to reflect non-hierarchically on the treatment of patients and on the hospital's operational modalities by creating a locus for exchanges and collegial decision making. These preparations also led to giving greater consideration to results of exit surveys and to committing procedures to paper, and were a key opportunity for introducing a continuous quality program.


Monodisperse NaYbF4:Tm3+/NaGdF4 core/shell nanocrystals with near-infrared to near-infrared upconversion photoluminescence and magnetic resonance properties.
We report core/shell NaYbF(4):Tm(3+)/NaGdF(4) nanocrystals to be used as probes for bimodal near infrared to near infrared (NIR-to-NIR) upconversion photoluminescence (UCPL) and magnetic resonance (MR) imaging. The NaYbF(4):Tm(3+) nanocrystals were previously reported to produce the intense NIR-to-NIR UCPL peaked at ∼800 nm under excitation at ∼975 nm. We have found that the growth of a NaGdF(4) shell on surface of the NaYbF(4):Tm(3+) nanocrystals results in the increase in the intensity of UCPL of Tm(3+) ions by about 3 times. Unlike biexponential PL decay of NaYbF(4):Tm(3+) nanocrystals, the PL decay of NaYbF(4):Tm(3+)/NaGdF(4) core/shell nanocrystals is single exponential and of longer lifetime due to the suppression of surface quenching effects for Tm(3+) PL. The growth of a NaGdF(4) shell on surface of the NaYbF(4):Tm(3+) nanocrystals also provides high MR relaxivity from paramagnetic Gd(3+) ions contained in the shell. The T1-weighted MR signal of the (NaYbF(4):2% Tm(3+))/NaGdF(4) nanoparticles was measured to be about 2.6 mM(-1)s(-1). Due to the combined presence of efficient optical and MR imaging capabilities, nanoprobes based on NaYbF(4):Tm(3+)/NaGdF(4) fluoride nanophosphors can be considered as a promising platform for simultaneous bimodal PL and MR bioimaging.


Social information use and social information waste.
Social information is immensely valuable. Yet we waste it. The information we get from observing other humans and from communicating with them is a cheap and reliable informational resource. It is considered the backbone of human cultural evolution. Theories and models focused on the evolution of social learning show the great adaptive benefits of evolving cognitive tools to process it. In spite of this, human adults in the experimental literature use social information quite inefficiently: they do not take it sufficiently into account. A comprehensive review of the literature on five experimental tasks documented 45 studies showing social information waste, and four studies showing social information being over-used. These studies cover 'egocentric discounting' phenomena as studied by social psychology, but also include experimental social learning studies. Social information waste means that human adults fail to give social information its optimal weight. Both proximal explanations and accounts derived from evolutionary theory leave crucial aspects of the phenomenon unaccounted for: egocentric discounting is a pervasive effect that no single unifying explanation fully captures. Cultural evolutionary theory's insistence on the power and benefits of social influence is to be balanced against this phenomenon. This article is part of the theme issue 'Foundations of cultural evolution'.


Role of sensory afferents in the myoelectric response to acute enteric inflammation in the rabbit.
The role of sensory afferents in inflammation-induced alterations in myoelectric activity in vivo was investigated in the rabbit small intestine. Isolated ileal loops were implanted with serosal electrodes and exposed to ricin or vehicle after pretreatment with 125 mg/kg of subcutaneous (125 mg over 3 days) or intraluminal (640 microM) capsaicin. After 5 h of myoelectric recording, the loops were prepared for histology and for ex vivo generation of eicosanoids. Capsaicin exacerbated mucosal damage after exposure to ricin but did not alter neutrophil infiltration. Subcutaneous capsaicin alone elevated slow-wave frequency and spike events and transiently suppressed the myoelectric response to ricin. In contrast, intraluminal capsaicin alone did not alter myoelectric activity but produced a sustained inhibition of the response to ricin. Eicosanoid production was unchanged by capsaicin alone. Intraluminal capsaicin blocked increases in leukotriene C4 and prostaglandin E2 during inflammation, an effect that paralleled its inhibition of myoelectric activity. Thus the contribution of sensory afferents to altered motility during acute ileitis involves the release of mucosal inflammatory mediators that influence neural control of smooth muscle.


[Epidural analgesia with clofelin in treating a pain syndrome].
The article presents a clinical experience with using epidural analgesia with Clofelin in 20 patients to arrest the pain syndrome developed at the postoperative period and resulting from atherosclerotic lesions of the lower extremity vessels. Control of the adequacy of anesthesia was made by registration of the H-reflex value. The investigations have shown that epidural analgesia with Clofelin is an effective measure to arrest the pain syndrome.


Mechanical models of sandfish locomotion reveal principles of high performance subsurface sand-swimming.
We integrate biological experiment, empirical theory, numerical simulation and a physical model to reveal principles of undulatory locomotion in granular media. High-speed X-ray imaging of the sandfish lizard, Scincus scincus, in 3 mm glass particles shows that it swims within the medium without using its limbs by propagating a single-period travelling sinusoidal wave down its body, resulting in a wave efficiency, η, the ratio of its average forward speed to the wave speed, of approximately 0.5. A resistive force theory (RFT) that balances granular thrust and drag forces along the body predicts η close to the observed value. We test this prediction against two other more detailed modelling approaches: a numerical model of the sandfish coupled to a discrete particle simulation of the granular medium, and an undulatory robot that swims within granular media. Using these models and analytical solutions of the RFT, we vary the ratio of undulation amplitude to wavelength (A/λ) and demonstrate an optimal condition for sand-swimming, which for a given A results from the competition between η and λ. The RFT, in agreement with the simulated and physical models, predicts that for a single-period sinusoidal wave, maximal speed occurs for A/λ ≈ 0.2, the same kinematics used by the sandfish.


ELISA on saliva samples for the detection of anti-hydatid cyst antibodies.
The detection of antibodies in saliva samples proved to be effective in the diagnosis of several microbial diseases. These antibodies were screened in saliva samples of patients with hydatid cysts. Anti-hydatid fluid antigen IgG and IgA antibodies were screened in saliva and sera of patients with hydatid cysts (n=37) as well as in healthy controls (n=30) using an in-house developed immunoenzymatic assay. Salivary anti-hydatid fluid antigen IgG showed a sensitivity of 86.5% and a specificity of 80%. A positive correlation was observed between anti-hydatid fluid antigen IgG in saliva and in serum (r = 0.364; p = 0.02). The detection of anti-hydatid fluid antigen IgG antibodies in saliva using ELISA promises to be interesting for the diagnosis of cystic echinococcosis.


Acute Hydrops with Total Corneal Edema in a Very Young Child with Keratoconus: The Youngest Age Reported Case.
To present the youngest age ever reported for acute corneal hydrops with total corneal edema in a child with advanced bilateral keratoconus. Patient presentation in ophthalmic clinic. The patient underwent various clinical tests and examinations including anterior segment optical coherence tomography (AS-OCT) and Scheimpflug corneal tomography. A 5-year-old girl presented with uncorrected distance visual acuity (UDVA) of 0.4 in the right eye and nonmeasurable UDVA associated with severe photophobia in her left eye of a 3-day duration. Intraocular pressure using the iCare tonometer was 14 and 5 mmHg in the right and left eyes, respectively. An old corneal hydrops scar and posterior subcapsular cataract (PSC) in the right eye and a total limbus to limbus corneal hydrops in the left eye were observed on slit-lamp examinations. Scheimpflug corneal tomography was possible in the right eye but, due to excessive irregularity and scaring, was not possible in the left eye. Corneal thinning and scarring were evident in the anterior segment optical coherence tomography in the right eye and very edematous cornea associated with stromal cleft and epithelial bullae in the left eye. A management plan consisting of topical hypertonic solution and ointment was started to reduce her symptoms. Acute corneal hydrops may be the presenting sign of keratoconus; however, extensive hydrops involving the total cornea area at a very young age is very rare and has not been previously reported in the literature.


The process for the activation of frog epidermis pro-tyrosinase.
1. Purified pro-tyrosinase from epidermis of the frog Rana esculenta ridibunda can be activated in vitro by several proteinases (trypsin, alpha-chymotrypsin, Pronase) and by light. 2. Both pro-tyrosinase and tyrosinase are composed of a single type of subunit having pI 7.2 and approximate molecular weights 68000 and 62000 respectively. A peptide of low molecular weight is released as a consequence of the proteolytic activation. Pro-tyrosinase and tyrosinase have different quaternary structures, the proenzyme being a dimer of Mr approx. 115000 and the enzyme a tetramer of Mr approx. 210 000. 3. The activation process was affected by several agents (L-3,4-dihydroxyphenylalanine, urea, formamide) that prevented, partially or totally, the activation of pro-tyrosinase. 4. The activation of pro-tyrosinase seems to be the result of a cleavage of the polypeptide chain that determines changes in tertiary or quaternary structure.


Validating eDNA measurements of the richness and abundance of anurans at a large scale.
In some situations, environmental DNA (eDNA) metabarcoding is a more accurate tool for measuring species richness of a taxon than conventional methods. Whether this tool can reliably estimate the abundance of a taxon remains unresolved. We examined the reliability of metabarcoding for measuring anuran diversity compared to a commonly used traditional line transect method (TLTM) through the replicate sampling of three visits across 71 waterbodies (ponds or reservoirs) in Liuheng, China. We also investigated the relative contributions of species-specific characteristics and the physiochemical properties of a waterbody on the relative read count across species and waterbodies. We found that eDNA metabarcoding had a higher detection probability for each of seven anuran species found in the sampling region than TLTM. Furthermore, the relative read count estimated by metabarcoding was positively correlated with the density or relative density of individuals identified with the TLTM across waterbodies for every species. Species-specific characteristics of anurans, such as density, relative density, body mass, biomass and relative biomass, accounted for substantial variations in the read count across species and waterbodies, while physiochemical factors, including pH, temperature, water volume, vegetation and elevation, had little effect on the read count. Our results based on robust sampling suggest that metabarcoding enables more reliable and efficient measurements of anuran occurrence at a large scale during a short-term survey (within 15 days) than that obtained by the TLTM, and offers an alternative tool for quantifications of anuran abundance. Density or biomass is better and more reliable indicator of anuran abundance associated with read count than relative density or relative biomass.


[Polymorphism of the genes IL-1RA and TNF-alpha in patients with gastritis and duodenal ulcer associated with Helicobacter pylori].
The goal of this work was to determine the rate of the polymorphism of the genes-antagonists of receptor IL-1 (IL-1RA) and TNF-alpha in patients with gastritis and duodenal ulcer associated with Helicobacter pylori. The receptors were tested for the clinical manifestation of the disease. A total of 126 patients with different gastroduodenal pathology and H. pylori in autopsy were tested. The results of this work demonstrated a correlation between the risk of duodenal ulcer and allele A of gene TNF-alpha in position 308 in the patients. The analysis of the gene-IL-1RA polymorphism demonstrated statistically significant difference between the patients in the frequency of the genotype 2/l. The results of this work showed that parallel typing of the genes of H. pylori in virulence was required for characterization of the bacteria-patient association. The correlation between the results of the typing with polymorphism of genes of cytokines in patient autopsy was also required.


EZH2 Expression in Retinoblastoma: A Potential Therapeutic Target.
The enhancer of zeste homolog 2 (EZH2) is a member of the polycomb repressive complex 2 (PRC2) and is important in cell-cycle regulation. Increased expression of EZH2 has been reported in retinoblastoma (RB). The aim of the study was to determine EZH2 expression, compare this with clinicopathological parameters in RB, and assess its relationship with tumor cell proliferation. Ninety-nine retrospective cases of enucleated RB were included in the present study. Expression of EZH2 and the marker of cell proliferation, Ki67, were investigated by immunohistochemistry. Among the 99 cases of RB in this study, EZH2 was found highly expressed (positive expression rate ≥70%) in 92 cases. EZH2 was expressed in tumor cells but absent in normal retinal tissues. The expression of EZH2 was positively linked to Ki67 expression (r = 0.65, p < 0.001). Elevated EZH2 expression was found in most RB cases, indicating that EZH2 could be a potential therapeutic target for RB.


Dangerous, depraved, and death-worthy: A meta-analysis of the correlates of perceived psychopathy in jury simulation studies.
Experimental research suggests that legal defendants described as psychopathic are generally, although not uniformly, judged more negatively and punitively. Understanding the correlates of perceived psychopathy, regardless of exposure to mental health evidence, is an important step towards clarifying divergent findings. We conducted a quantitative synthesis of ten juror simulation studies (combined N = 2,980) examining the meta-analytic association between perceived defendant psychopathy and various psychologically important and legally relevant outcomes. Perceiving someone as being more psychopathic was associated with viewing that defendant as more dangerous (r W = 0.31) and evil ( r W = 0.44). Moreover, perceptions of defendant psychopathy predicted greater support for more adverse consequences in terms of capital sentencing ( r W = 0.22) and sentence length ( r W = 0.27), although not perceived treatment amenability ( r W = 0.09). These findings highlight the importance of including ratings of perceived psychopathy in experimental designs to identify the circumstances under which psychopathy evidence might prejudicially impact case outcomes.


PiB Fails to Map Amyloid Deposits in Cerebral Cortex of Aged Dogs with Canine Cognitive Dysfunction.
Dogs with Canine Cognitive Dysfunction (CCD) accumulate amyloid beta (Aβ) in the brain. As the cognitive decline and neuropathology of these old dogs share features with Alzheimer's disease (AD), the relation between Aβ and cognitive decline in animal models of cognitive decline is of interest to the understanding of AD. However, the sensitivity of the biomarker Pittsburgh Compound B (PiB) to the presence of Aβ in humans and in other mammalian species is in doubt. To test the sensitivity and assess the distribution of Aβ in dog brain, we mapped the brains of dogs with signs of CCD (n = 16) and a control group (n = 4) of healthy dogs with radioactively labeled PiB ([(11)C]PiB). Structural magnetic resonance imaging brain scans were obtained from each dog. Tracer washout analysis yielded parametric maps of PiB retention in brain. In the CCD group, dogs had significant retention of [(11)C]PiB in the cerebellum, compared to the cerebral cortex. Retention in the cerebellum is at variance with evidence from brains of humans with AD. To confirm the lack of sensitivity, we stained two dog brains with the immunohistochemical marker 6E10, which is sensitive to the presence of both Aβ and Aβ precursor protein (AβPP). The 6E10 stain revealed intracellular material positive for Aβ or AβPP, or both, in Purkinje cells. The brains of the two groups of dogs did not have significantly different patterns of [(11)C]PiB binding, suggesting that the material detected with 6E10 is AβPP rather than Aβ. As the comparison with the histological images revealed no correlation between the [(11)C]PiB and Aβ and AβPP deposits in post-mortem brain, the marked intracellular staining implies intracellular involvement of amyloid processing in the dog brain. We conclude that PET maps of [(11)C]PiB retention in brain of dogs with CCD fundamentally differ from the images obtained in most humans with AD.


Exploring young adults' e-cigarette use behavior during COVID-19.
Changes in daily life related to COVID-19 have impacted e-cigarette use, particularly in young adults. This cross-sectional mixed-methods study explored young adults' perceptions regarding how COVID-19 influenced their e-cigarette use. We analyzed Fall 2020 survey data from 726 past 6-month e-cigarette users (mean age=24.15 years, 51.1% female, 35.5% sexual minority, 4.4% Black, 10.2% Asian, 12.1% Hispanic) and Spring 2021 semi-structured interview data among a subset of 40 participants (mean age=26.30 years, 35.0% female, 45.0% sexual minority, 5.0% Black, 22.5% Asian, 12.5% Hispanic). Participants were drawn from 6 metropolitan statistical areas with varied tobacco and cannabis legislative contexts. Among survey participants, 44.4% also smoked cigarettes, 54.0% other tobacco products, and 60.1% used cannabis. They reported various changes in their daily lives, including changes in the nature and/or status of employment (e.g. 15.3% were laid off, 72.8% experienced household income loss). Regarding changes in e-cigarette use since COVID-19, 22.6% tried to cut down and 16.0% tried to quit. Interview participants commonly indicated that they increased their use due to stress, boredom, changes in accessibility, and/or changes to daily environment that made e-cigarette use more feasible. Results highlight the importance of promoting opportunities for young adults to build relationships to decrease stress, foster a sense of belonging, and increase quality of life (e.g. increasing the accessibility to mental health and social support services, intentionally engaging young adults in pandemic-appropriate community-building and extracurricular activities). This research may help to inform future e-cigarette cessation interventions that consider the unique challenges of societal stressors, such as pandemics.


Analysis of the mechanism of d-amphetamine-and apomorphine-induced changes of ascorbic acid catabolism in discrete brain areas of the rat.
Ascorbic acid (AA) levels and dehydroascorbic acid/ascorbic acid (DHAA/AA) ratios were determined in hypothalamus, striatum, and remaining brain of male Wistar rats, after single or repeated injections of d-amphetamine (1.8 mg/kg/day s.c.), apomorphine (1.0 mg/kg/day s.c.), and/or haloperidol (0.1 mg/kg/day i.p.). Major changes were observed in hypothalamus, in which all drugs significantly increased DHAA/AA ratio. Apomorphine and haloperidol consistently decreased AA level as well. The DHAA/AA ratio increase was observed also when apomorphine and d-amphetamine were associated with haloperidol. In striatum, apomorphine (single) and d-amphetamine (repeated) injections increased DHAA/AA ratios; such AA oxidation increase was inhibited by haloperidol; the increases (d-amphetamine) or decrease (apomorphine) of AA levels were also inhibited by haloperidol; haloperidol alone did not modify DHAA/AA ratio and induced minor changes of striatal AA level. In remaining brain, apomorphine (single) and d-amphetamine (repeated) treatment increased DHAA/AA ratio; such increase was observed also when haloperidol was associated with apomorphine or d-amphetamine; moreover, haloperidol by itself increased AA oxidation, although to a lesser extent than it did in hypothalamus. It is concluded that AA catabolism can be activated in the rat striatum by a dopaminergic mechanism; in hypothalamus and in remaining brain, the AA catabolism activation appears rather to be a non-specific effect of the pharmacological manipulation.


Misdiagnosed hypomanic symptoms in patients with treatment-resistant major depressive disorder in Italy: results from the improve study.
Undiagnosed and therefore inadequately treated hypomanic symptoms may be a leading cause of drug resistance in depression diagnosed as unipolar (major depressive disorder, MDD). The purpose of the IMPROVE study was to identify the rate of misdiagnoses in patients with treatment-resistant MDD by screening for the presence of previous hypomanic episodes, and to study the characteristics of those patients with a positive history of hypomania. Patients attending 29 psychiatric units throughout Italy with a diagnosis of MDD who were resistant to anti-depressant treatment were included in this multicentre, observational single visit study. The Hypomania Checklist 32 (HCL-32) was administered to detect underlying bipolarity. Among the 466 enrolled patients, 256 (57.40%) were positive at screening for a previous hypomanic episode (HCL-32 ≥12), therefore suggesting a misdiagnosis. These patients scored higher than those with a negative history in both the "active/elated hypomania" (11.27±3.11 vs 3.57±3.05; P<0.0001) and "irritable/risk-taking hypomania" (2.87±2.03 vs 2.06±1.73; P<0.001) HCL-32 sub-scales. Patients with a positive history of hypomania were younger, had a higher number of previous depressive episodes and a higher frequency of comorbid conditions compared to those with a negative history. This study suggests that screening for hypomania in MDD-resistant patients facilitates identification of a notable proportion of undiagnosed cases of bipolar spectrum disorder. Patients with a positive history of hypomania at screening had a demographic/clinical bipolar-like profile that included young age, higher number of previous depressive episodes and higher frequency of comorbid conditions. They also had both higher active and irritable hypomania symptom scores.


Methyl-coenzyme M formation in methanogenic archaea. Involvement of zinc in coenzyme M activation.
Methyl-coenzyme M (2-methylthioethane sulfonate) is the key intermediate of methane formation in methanogenic archaea. It is generated from coenzyme M (2-mercaptoethane sulfonate) in methyl transfer reactions catalyzed by proteins containing zinc. Here, we report that, for methyltransferase MtaA from Methanosarcina barkeri, the zinc is involved in coenzyme M activation. For the experiments an inactive MtaA apoprotein was obtained by heterologous overproduction in Escherichia coli grown in the presence of 2 mM EDTA. The apoprotein was found to react with zinc or cobalt to the fully active holoenzyme. Appoximately 1 mol of transition metal was bound per mol of protein. Upon incubation of the holoenzyme with coenzyme M approximately 1 mol of proton was released per mol of zinc or cobalt. Protons were not released upon incubation of the apoprotein with coenzyme M or of the holoprotein with other thiol compounds or with methyl-coenzyme M. The findings are interpreted as indicating that the role of the transition metal in MtaA is to lower the microscopic pKa of the thiol group of coenzyme M by coordination to the zinc, and thus to increase its nucleophilicity for methyl group attack. The pKZn2+ of MtaA was re-determined and found to be > 15 and not 9.6 as previously reported by us.


Restrictions in cell cycle progression of adult vestibular supporting cells in response to ectopic cyclin D1 expression.
Sensory hair cells and supporting cells of the mammalian inner ear are quiescent cells, which do not regenerate. In contrast, non-mammalian supporting cells have the ability to re-enter the cell cycle and produce replacement hair cells. Earlier studies have demonstrated cyclin D1 expression in the developing mouse supporting cells and its downregulation along maturation. In explant cultures of the mouse utricle, we have here focused on the cell cycle control mechanisms and proliferative potential of adult supporting cells. These cells were forced into the cell cycle through adenoviral-mediated cyclin D1 overexpression. Ectopic cyclin D1 triggered robust cell cycle re-entry of supporting cells, accompanied by changes in p27(Kip1) and p21(Cip1) expressions. Main part of cell cycle reactivated supporting cells were DNA damaged and arrested at the G2/M boundary. Only small numbers of mitotic supporting cells and rare cells with signs of two successive replications were found. Ectopic cyclin D1-triggered cell cycle reactivation did not lead to hyperplasia of the sensory epithelium. In addition, a part of ectopic cyclin D1 was sequestered in the cytoplasm, reflecting its ineffective nuclear import. Combined, our data reveal intrinsic barriers that limit proliferative capacity of utricular supporting cells.


A new technique to evaluate the impact of running on knee cartilage deformation by region.
When measuring changes in knee cartilage thickness in vivo after loading, mean values may not reflect local changes. The objectives of this investigation were: (1) use statistical parametric mapping (SPM) to determine regional deformation patterns of tibiofemoral cartilage in response to running; (2) quantify regional differences in cartilage thickness between males and females; and (3) explore the influence of sex on deformation. Asymptomatic males (n = 15) and females (n = 15) had MRI imaging of their right knee before and after 15 min of treadmill running. Medial and lateral tibial, and medial and lateral weight-bearing femoral cartilage were segmented. SPM was completed on cartilage thickness maps to test the main effects of Running and Sex, and their interaction. F-statistic maps were thresholded; clusters above this threshold indicated significant differences. Deformation was observed in all four compartments; the lateral tibia had the largest area of deformation (p < 0.0001). Thickness differences between sexes were observed in all four compartments, showing females have thinner cartilage (p ≤ 0.009). The lateral tibia had small clusters indicating an interaction of sex on deformation (p ≤ 0.012). SPM identified detailed spatial information on tibiofemoral cartilage thickness differences observed after running, and between sexes and their interaction.


The speed-curvature power law in Drosophila larval locomotion.
We report the discovery that the locomotor trajectories of Drosophila larvae follow the power-law relationship between speed and curvature previously found in the movements of human and non-human primates. Using high-resolution behavioural tracking in controlled but naturalistic sensory environments, we tested the law in maggots tracing different trajectory types, from reaching-like movements to scribbles. For most but not all flies, we found that the law holds robustly, with an exponent close to three-quarters rather than to the usual two-thirds found in almost all human situations, suggesting dynamic effects adding on purely kinematic constraints. There are different hypotheses for the origin of the law in primates, one invoking cortical computations, another viscoelastic muscle properties coupled with central pattern generators. Our findings are consistent with the latter view and demonstrate that the law is possible in animals with nervous systems orders of magnitude simpler than in primates. Scaling laws might exist because natural selection favours processes that remain behaviourally efficient across a wide range of neural and body architectures in distantly related species.